Drug metabolism is an important factor in development of drugs. Especially, deficiency of drug metabolism in a body causes severe side effects such as exaggerated pharmacological response and toxic response.
Previous studies have revealed that cytochrome P-450 isozymes are responsible for drug metabolism, and oxidation by P-450 isozymes is a common aspect of the overall clearance of drugs. Further studies have revealed that genetic polymorphism of cytochrome P-450 isozymes underlies a wide spectrum of substrates specificity in drug oxidation. In certain cases, genetic mutation and/or deletion of one critical isozyme gene results in a significant alteration of a phenotype projected on substrate specificity. More specifically for example, defect of CYP2D6 gene is known to be responsible for poor metabolism of debrisoquine or spateteine (for example, H. J. Dengler et al., Arzenmittel-Forshung, Vol. 27, pp. 1836-, 1977). In addition, it has been reported that CYP2D6 oxidizes more than 30 drugs (for example, M. Eichelbaum et al., Pharmacol. Ther., Vol. 46, pp. 377-, 1990). Thus, those who have homozygous alteration in this recessive gene, are so-called "poor metabolizers (PMs)" and may suffer from severe side effects due to poor metabolism of drugs (for example, see M. Eichelbaum et al., Pharmacol. Ther., Vol. 46, pp. 377-, 1990). Such genetic alterations occur at rates of from 1 to 30% in different ethnic populations (for example, L. M. Distlerath et al., J. Biol. Chem., Vol. 260, pp. 9057-, 1985).
Previously, drug metabolism was evaluated by adding quinidine to a buffer solution of a substrate and microsomes (F. Peter et al., Molecular Pharmacology, Vol. 30, pp. 287-295, 1986). In this assay, quinidine is a strong competitive inhibitor of CYP2D6, and thus the contribution of CYP2D6 to the oxidative metabolism can be evaluated. However, quinidine is reported to inhibit other P-450 isozymes such as CYP3A4 (for example, F. P. Guengerich et al., Molecular Pharmacology, Vol. 30, pp. 287-, 1986), thereby preventing accurate evaluation of contribution of CYP2D6 in drug metabolism.
Therefore, it would be desirable if there were provided an improved method for accurately evaluating the susceptibility of a sample compound to metabolism with respect to a specific P-450 isozyme.